scholarly journals Characterisation and antibiotic susceptibility profile of Clostridioides (Clostridium) difficile isolated from chicken carcasses

2020 ◽  
Vol 64 (3) ◽  
pp. 407-412 ◽  
Author(s):  
Enver Baris Bingol ◽  
Hamparsun Hampikyan ◽  
Karlo Muratoglu ◽  
Esra Akkaya ◽  
Omer Cetin ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Antibiotic Susceptibility ◽  
Inhibitory Concentration ◽  
Toxin Production ◽  
Ribotype 027 ◽  
Food Borne ◽  
Chicken Carcass ◽  
Microbiological Methods ◽  
Common Causes ◽  
Amoxicillin Clavulanic Acid

AbstractIntroductionClostridioides (Clostridium) difficile is a Gram+, anaerobic, spore-forming, rod-shaped bacterium that can produce toxins, and it is mainly because its virulence is attributed. The objective of this study was to evaluate the presence of C. difficile and hyper virulent ribotypes in chicken carcasses and the antibiotic susceptibility of isolated strains.Material and MethodsC. difficile was isolated from chicken carcasses by microbiological methods, its ribotypes were identified by means of PCR, the toxin production ability was defined by ELISA, and the susceptibility of the isolates to selected antibiotics was determined by minimum inhibitory concentration evaluator strips.ResultsThe bacterium was isolated from 69 out of 185 (37.3%) examined chicken carcass samples, and six out of the 69 (8.7%) isolates were identified as ribotype 027. All isolates were susceptible to amoxicillin-clavulanic acid (100.0%), vancomycin (97.1%), metronidazole (88.4%), and tetracycline (95.7%), whereas they were resistant to cefotaxime (97.1%) and imipenem (89.9%).ConclusionThe results of this study demonstrate the presence of toxigenic C. difficile isolates such as ribotype 027 (one of the most common causes of C. difficile infection in humans) in chicken carcasses. Although there is no case for stating that C. difficile is a food-borne pathogen, the presence of C. difficile in chicken may be considered to be a potential risk to consumers.

Antimicrobial Susceptibility Pattern of Staphylococcus Species from Clinical Samples

2020 ◽  
Author(s):  
Muhammad Ali
Keyword(s):  
Clavulanic Acid ◽  
Antibiotic Susceptibility ◽  
Vaginal Swab ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Susceptibility Pattern ◽  
Mannitol Salt Agar ◽  
Agar Disc ◽  
Microbiological Methods ◽  
Amoxicillin Clavulanic Acid

The research was aimed to evaluate the antibiotic susceptibility pattern of Staphylococcus species from clinical samples obtained from some hospitals in Kano metropolis, Nigeria. The ear swab, high vaginal swab (HVS), wound swab and urine samples from the patients attending the hospitals were collected and inoculated onto the surface of freshly prepared Nutrient agar for bacterial isolation. The bacteria isolated were identified by conventional microbiological methods namely; Gram staining, biochemical test (such as catalase, coagulase, and DNase test), mannitol salt agar and heamolysis test. The isolates were subjected to antibiotic susceptibility testing using the agar disc diffusion method. The result showed that S. aureus was highly susceptible to Ciprofloxacin 105 (68.63%), Gentamicin 102 (66.67%), Levofloxacin 95 (62.08%) and Amikacin 90 (58.82%), S. epidermidis was highly susceptible to Gentamicin 13 (61.90%), Levofloxacin 12 (57.14%) and Nitrofurantoin 11 (52.38%) while S. saprophyticus was highly susceptible to Cefoxitin 7 (77.78%), Gentamicin 6 (66.67%) and Nitrofurantoin 5 (55.56%). On the other hand, S. aureus was highly resistant to Cefuroxime 153 (100%), Ceftazidime 150 (98.04%), Amoxicillin/clavulanic acid 120 (78.43%) and Cloxacillin 111 (72.55%), S. epidermidis was highly resistant to Ceftazidime 20 (95.24%), Cloxacillin 19 (90.48%) then Cefoxitin, Erythromycin and Amoxicillin/clavulanic acid with 15 (71.43%) both. S. saprophyticus was highly resistant to Cefepime 9 (100%), Cloxacillin 8 (88.89%), Ceftazidime 7 (77.78%), Imipenem and Erythromycin with 6 (66.67%) respectively. There is a statistical difference in the sensitivity of the isolates against the antibiotics used at p<0.05. It is concluded that Staphylococcus species develop resistance to some classes of antibiotics.

In vitro antibiotic susceptibility profile of Clostridium difficile excluding PCR ribotype 027 outbreak strain in Hungary

Anaerobe ◽  
2014 ◽  
Vol 30 ◽  
pp. 41-44 ◽  
Author(s):  
Gabriella Terhes ◽  
Akiko Maruyama ◽  
Krisztina Latkóczy ◽  
Lenke Szikra ◽  
Marianne Konkoly-Thege ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Antibiotic Susceptibility ◽  
Outbreak Strain ◽  
Ribotype 027 ◽  
Pcr Ribotype 027

scholarly journals Effects of Exposure of Clostridium difficile PCR Ribotypes 027 and 001 to Fluoroquinolones in a Human Gut Model

2008 ◽  
Vol 53 (2) ◽  
pp. 412-420 ◽  
Author(s):  
Katie Saxton ◽  
Simon D. Baines ◽  
Jane Freeman ◽  
Rachael O'Connor ◽  
Mark H. Wilcox
Keyword(s):  
Clostridium Difficile ◽  
Gut Microbiota ◽  
Spore Germination ◽  
Toxin Production ◽  
Human Gut ◽  
Ribotype 027 ◽  
Resistant Mutants ◽  
High Level ◽  
Pcr Ribotype 027

ABSTRACT The incidence of Clostridium difficile infection is increasing, with reports implicating fluoroquinolone use. A three-stage chemostat gut model was used to study the effects of three fluoroquinolones (ciprofloxacin, levofloxacin, and moxifloxacin) on the gut microbiota and two epidemic C. difficile strains, strains of PCR ribotypes 027 and 001, in separate experiments. C. difficile total viable counts, spore counts, and cytotoxin titers were determined. The emergence of C. difficile isolates with reduced antibiotic susceptibility was monitored with fluoroquinolone-containing medium, and molecular analysis of the quinolone resistance-determining region was performed. C. difficile spores were quiescent in the absence of fluoroquinolones. Instillation of each fluoroquinolone led to C. difficile spore germination and high-level cytotoxin production. High-level toxin production occurred after detectable spore germination in all experiments except those with C. difficile PCR ribotype 027 and moxifloxacin, in which marked cytotoxin production preceded detectable germination, which coincided with isolate recovery on fluoroquinolone-containing medium. Three C. difficile PCR ribotype 027 isolates and one C. difficile PCR ribotype 001 isolate from fluoroquinolone-containing medium exhibited elevated MICs (80 to ≥180 mg/liter) and possessed mutations in gyrA or gyrB. These in vitro results suggest that all fluoroquinolones have the propensity to induce C. difficile infection, regardless of their antianaerobe activities. Resistant mutants were seen only following moxifloxacin exposure.

Toxin A and B genes expression of Clostridium difficile in the sub-minimum inhibitory concentration of clindamycin, vancomycin and in combination with ceftazidime

2020 ◽  
Author(s):  
Mohammad Moradi ◽  
Shahla Mansouri ◽  
Nouzar Nakhaee ◽  
Farhad Sarafzadeh ◽  
Ebrahim Rezazadeh Zarandi
Keyword(s):  
Gene Expression ◽  
Clostridium Difficile ◽  
Inhibitory Concentration ◽  
Toxin Production ◽  
Toxin Gene ◽  
Transcription Level ◽  
The Third ◽  
Genes Expression ◽  
Antibiotic Associated Diarrhea ◽  
Toxin Gene Expression

Background and Objectives: Antibiotics prescribed for infections have diverse effects on microbiota and the pathogen Clostridium difficile (C. difficile) as the most important antibiotic-associated diarrhea. This study aims to determine the gene expression of toxins A and B at the transcription level in the sub-MIC of vancomycin (VAN), clindamycin (CLI), and cef- tazidime (CAZ) alone and in combination. Materials and Methods: The MIC and fractional inhibitory concentration (FIC) of two C. difficile samples (a clinical isolate and ATCC 9689) were determined by microdilution and checkerboard microdilution methods, respectively. The total RNA was extracted from the medium inoculated with ~106  CFU/mL of fresh bacteria in the pre-reduced medium containing  ½ MIC of antibiotics alone and ½ FIC of antibiotics in combination. Real-time PCR was performed by sybrGreen methods in triplicate, and the data were analyzed by the comparative ∆∆CT  method. Results: All antibiotics except CAZ (alone and in combination) decreased the gene expression of toxins A and B within 24 hours. VAN and CLI reduced toxin gene expression within 24 and 48 hours. However, CAZ alone and in combination with VAN as well as CLI increased the gene expression of toxins A and B. Conclusion: The results confirmed toxin gene transcription and toxin production are associated with the type of isolates and antibiotics, as well as the combined form of antibiotics. This could be the reason which can explain the occurrence of C. difficile infection among patients who were treated with the third generation of cephalosporins alone and in combination with another antibiotic in the form of combinational therapy.

scholarly journals Comparison of toxin and spore production in clinically relevant strains of Clostridium difficile

Microbiology ◽  
2011 ◽  
Vol 157 (5) ◽  
pp. 1343-1353 ◽  
Author(s):  
Prerna Vohra ◽  
Ian R. Poxton
Keyword(s):  
Clostridium Difficile ◽  
Stationary Phases ◽  
Toxin Production ◽  
Spore Production ◽  
Phenotypic Traits ◽  
Ribotype 027 ◽  
Repressive Effect ◽  
Changing Trend ◽  
Different Strains

Clostridium difficile is a major cause of nosocomial diarrhoea. The toxins that it produces (TcdA and TcdB) are responsible for the characteristic pathology of C. difficile infection (CDI), while its spores persist in the environment, causing its widespread transmission. Many different strains of C. difficile exist worldwide and the epidemiology of the strains is ever-changing: in Scotland, PCR ribotype 012 was once prevalent, but currently ribotypes 106, 001 and 027 are endemic. This study aimed to identify the differences among these ribotypes with respect to their growth, and toxin and spore production in vitro. It was observed that the hypervirulent ribotype 027 produces significantly more toxin than the other ribotypes in the exponential and stationary phases of growth. Further, the endemic strains produce significantly more toxins and spores than ribotype 012. Of note was the observation that tcdC expression did not decrease into the stationary phase of growth, implying that it may have a modulatory rather than repressive effect on toxin production. Further, the increased expression of tcdE in ribotype 027 suggests its importance in the release of the toxins. It can thus be concluded that several genotypic and phenotypic traits might synergistically contribute to the hypervirulence of ribotype 027. These observations might suggest a changing trend towards increased virulence in the strains currently responsible for CDI.

scholarly journals The In Vitro Antibiotic Susceptibility of Malaysian Isolates ofBurkholderia pseudomallei

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Norazah Ahmad ◽  
Rohaidah Hashim ◽  
Azura Mohd Noor
Keyword(s):  
Antibiotic Susceptibility ◽  
Burkholderia Pseudomallei ◽  
Inhibitory Concentration ◽  
Test Method ◽  
Amoxicillin Clavulanic Acid ◽  
Antibiotic Susceptibility Patterns ◽  
Potential Alternative ◽  
Ciprofloxacin Resistance ◽  
Susceptibility Patterns

Acute melioidosis may present as localised or septicaemic infections and can be fatal if left untreated.Burkholderia pseudomalleiresistant to antibiotics used for the treatment of melioidosis had been reported. The aim of this study was to determine the in vitro antibiotic susceptibility patterns ofBurkholderia pseudomalleiisolated in Malaysia to a panel of antibiotics used for the treatment of melioidosis and also to potential alternative antibiotics such as tigecycline, ampicillin/sulbactam, and piperacillin/tazobactam. A total of 170Burkholderia pseudomalleiisolates were subjected to minimum inhibitory concentration determination usingE-test method to eleven antibiotics. All isolates were sensitive to meropenem and piperacillin/tazobactam. For ceftazidime, imipenem, amoxicillin/clavulanic acid, and doxycycline resistance was observed in 1 isolate (0.6%) for each of the antibiotics. Trimethoprim/sulfamethoxazole resistance was observed in 17 (10%) isolates. For other antibiotics, ampicillin/sulbactam, chloramphenicol, tigecycline, and ciprofloxacin resistance were observed in 1 (0.6%), 6 (3.5%), 60 (35.3%) and 98 (57.7%) isolates respectively. One isolate B170/06 exhibited resistance to 4 antibiotics, namely, ciprofloxacin, chloramphenicol, trimethoprim/sulfamethoxazole, and tigecycline. In conclusion, the Malaysian isolates were highly susceptible to the current antibiotics used in the treatment of melioidosis in Malaysia. Multiple resistances to the antibiotics used in the maintenance therapy are the cause for a concern.

scholarly journals Precise Manipulation of the Clostridium difficile Chromosome Reveals a Lack of Association between thetcdCGenotype and Toxin Production

2012 ◽  
Vol 78 (13) ◽  
pp. 4683-4690 ◽  
Author(s):  
Stephen T. Cartman ◽  
Michelle L. Kelly ◽  
Daniela Heeg ◽  
John T. Heap ◽  
Nigel P. Minton
Keyword(s):  
Clostridium Difficile ◽  
Diarrheal Disease ◽  
Genetic Manipulation ◽  
Toxin Production ◽  
Negative Regulator ◽  
Toxin B ◽  
Content Type ◽  
Ribotype 027 ◽  
High Level ◽  
Pcr Ribotype 027

ABSTRACTClostridium difficilecauses a potentially fatal diarrheal disease through the production of its principal virulence factors, toxin A and toxin B. ThetcdCgene is thought to encode a negative regulator of toxin production. Therefore, increased toxin production, and hence increased virulence, is often inferred in strains with an aberranttcdCgenotype. This report describes the first allele exchange system for precise genetic manipulation ofC. difficile, using thecodAgene ofEscherichia colias a heterologous counterselection marker. It was used to systematically restore the Δ117 frameshift mutation and the 18-nucleotide deletion that occur naturally in thetcdCgene ofC. difficileR20291 (PCR ribotype 027). In addition, the naturally intacttcdCgene ofC. difficile630 (PCR ribotype 012) was deleted and then subsequently restored with a silent nucleotide substitution, or “watermark,” so the resulting strain was distinguishable from the wild type. Intriguingly, there was no association between thetcdCgenotype and toxin production in eitherC. difficileR20291 orC. difficile630. Therefore, an aberranttcdCgenotype does not provide a broadly applicable rationale for the perceived notion that PCR ribotype 027 strains are “high-level” toxin producers. This may well explain why several studies have reported that an aberranttcdCgene does not predict increased toxin production or, indeed, increased virulence.

scholarly journals Spectral and Antibiotic Susceptibility of Pathogens Isolated from Saudi Patients with Diabetic Foot Infections

2021 ◽  
Vol 12 (1) ◽  
pp. 16-20
Author(s):  
Samiah Hamad S Al-Mijalli
Keyword(s):  
Staphylococcus Aureus ◽  
Diabetic Foot ◽  
Antibiotic Susceptibility ◽  
Empirical Therapy ◽  
Multidrug Resistant ◽  
Foot Ulcers ◽  
Sensitivity Testing ◽  
Cross Sectional ◽  
Diabetic Foot Infections ◽  
Microbiological Methods

Diabetic foot infections (DFIs) are a significant health issue and a common complication among patients with diabetes. To develop antibiotic therapy for these high-risk patients, the current study evaluates the scope of DFIs and identifies the causing microbes. It also measures spectrum and antibiotic susceptibility of the pathogens isolated from adults with DFIs in Saudi Arabia. To achieve the study objectives, a cross-sectional study was implemented and the baseline characteristics for 44 patients with DFIs were defined. Optimal aerobic and anaerobic microbiological techniques were utilized to culture specimens isolated from infected foot ulcers. The standard microbiological methods were employed to identify the bacterial isolates and antibiotic susceptibility testing was conducted following the procedures of the Clinical and Laboratory Standards Institute (CLSI). Results showed that 12 microorganisms were isolated from the participants’ diabetic foot ulcers. Staphylococcus Aureus was ranked first because it appeared in 29 (65.9%) cases. Streptococcus Agalactiae was ranked second and multi-microbial infections were also found. Most of the organisms were susceptible to Vancomycin, Ciprofloxacin, and Cefalexin, but they were resistant to Methicillin, Gentamicin, and Ampicillin antibiotics. Staphylococcus Aureus was most sensitive to Ciprofloxacin, while it was resistant to Methicillin. About 10% of the isolates were multidrug-resistant. The study concludes that while Vancomycin should be used empirically for Gram-positive isolates, Ciprofloxacin can be taken into consideration for most of the Gram-negatives aerobes. Based on including various microorganisms and the advent of multidrug-resistant strains, proper culture and sensitivity testing are necessary prior to the empirical therapy.

scholarly journals Antibiotic use and other risk factors at hospital level for outbreaks with Clostridium difficile PCR ribotype 027

2008 ◽  
Vol 57 (6) ◽  
pp. 709-716 ◽  
Author(s):  
T. I. I. van der Kooi ◽  
M. Koningstein ◽  
A. Lindemans ◽  
D. W. Notermans ◽  
E. Kuijper ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Antibiotic Use ◽  
Study Groups ◽  
Ribotype 027 ◽  
Defined Daily Doses ◽  
Hygiene Measures ◽  
Epidemic Period ◽  
Group A ◽  
Group B ◽  
The Relationship

The first Dutch outbreak due to Clostridium difficile ribotype 027 was observed in mid-2005; by the end of that year, eight hospitals were affected. To study the relationship between hospital-wide antibiotic use and the incidence of 027-linked C. difficile-associated disease (CDAD) three study groups were made: group A, all eight hospitals with an 027-associated epidemic; group B, five of a total of six hospitals with occasional 027 cases, without an increase in CDAD; and group C, ten randomly selected hospitals with no reported 027 epidemics or isolated 027 cases. Quarterly data on CDAD incidences, hygiene measures and the use of fluoroquinolones, second- and third-generation cephalosporins, extended-spectrum penicillins, penicillins with beta-lactamase inhibitors, carbapenems, lincomycins and macrolides were collected for 2004 and 2005, and divided into pre-epidemic and epidemic periods. Using a multilevel Poisson regression analysis, CDAD incidence was linked to antibiotic use in the previous quarter and to certain hygiene measures. In the pre-epidemic period, the total use of the studied antibiotics was comparable between affected and unaffected hospitals. Higher use of second-generation cephalosporins, macrolides and all of the studied antibiotics were independently associated with a small increase in CDAD incidence [relative risk (95 % confidence interval): 1.14 per increase of 100 defined daily doses per 10 000 bed days (1.06–1.23), 1.10 (1.01–1.19) and 1.02 (1.01–1.03), respectively]. However the effect was too small to predict which hospitals might be more prone to 027-associated outbreaks.

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